N-cadherin Antibody (8C11): sc-53488

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	-Mouse (1:0000) -Use clean forceps to gently handle the blot from the corner without creasing the membrane. Do not write on the blot with pen or marker, as the ink can fluoresce and cause background.

Protocol tips
-Mouse (1:0000) -Use clean forceps to gently handle the blot from the corner without creasing the membrane. Do not write on the blot with pen or marker, as the ink can fluoresce and cause background.

Publication protocol

Cells from all experimental groups were collected using a cell scraper and proteins were extracted by cell lysis buffer. Total protein was extracted from human NPC lines and liver tissues using radioimmunoprecipitation assay buffer (Sigma-Aldrich; Merck Millipore, Darmsdadt, Germany). Protein concentration was assessed with a Bradford assay. Equal quantities of cell and tissue lysates (30 µg protein) were separated by 10% SDS-PAGE, transferred onto a polyvinylidene difluoride membrane (EMD Millipore, Billerica, MA, USA) and blocked with 5% skimmed milk powder for 1 h at room temperature. Membranes were then incubated with p300 antibody (catalog no. sc-584; 1:250 dilution; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) against target proteins at 4°C, followed by washing with TBS and Tween-20 (TBST) overnight. Membranes were then incubated with the horseradish peroxidase-labeled secondary antibody (1:1,000 dilution; YN-16; Forevergen Biosciences) for 1 h at 37°C, and then washed with TBST. The immunoreactive signals were detected with enhanced chemiluminescence (Forevergen Biosciences). GAPDH was used as a loading control.

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Manufacturer protocol

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